Site-directed mutagenesis of UbiA to promote menaquinone biosynthesis in Elizabethkingia meningoseptica |
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| Affiliation: | 1. College of Biochemical Engineering, Anhui Polytechnic University, 241000, Wuhu, China;2. Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, 23298, Richmond, VA, USA;3. School of Biological Science and Food Engineering, Chuzhou University, 239000, Chuzhou, China;1. CNRS, LAAS, 7 avenue du colonel Roche, F-31400 Toulouse, France;2. Univ de Toulouse, LAAS, F-31400 Toulouse, France;3. CNRS, ITAV-USR3505, Toulouse, France;4. Univ de Toulouse, ITAV-USR3505, Toulouse, France;5. Univ de Toulouse, INSA, LAAS, F-31400 Toulouse, France;1. Institut für Statistik und Operations Research, Universität Graz, Universitätsstraße 15, A-8010 Graz, Austria;2. Department of Industrial Engineering and Operations Research, University of California, Berkeley, CA 94720, USA;3. Department of Mathematical Sciences, University of Greenwich, Old Royal Naval College, Park Row, Greenwich, London SE10 9LS, UK;1. Regional Medical Research Centre (ICMR), Department of Health Research, Nehru Nagar, Belagavi 590 010, Karnataka, India;2. Department of Biotechnology, Manipal Institute of Technology, Manipal University, Manipal, Karnataka, India;1. Laboratory of Biochemistry, Gifu Pharmaceutical University, Gifu, Japan;2. Monash Institute of Pharmaceutical Sciences, Monash University, Victoria, Australia;3. Faculty of Engineering, Gifu University, Gifu, Japan |
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| Abstract: | To increase the menaquinone (MK) content of an Elizabethkingia meningoseptica, site-directed mutagenesis was generated to suppress 4-hydroxybenzoate octaprenyl transferase (UbiA) activity and subsequently blocked the ubiquinone (UQ) biosynthesis pathway. Fourteen conserved residues except L174 and G211 were mutated to analyze the effect of site-directed mutagenesis. The expression of UbiA in twelve mutants was decreased in both mRNA and protein levels, which resulted in the decrease of UQ concentration. Based on MenA expression level, 12 mutants were divided into two groups. Second group such as N72A, D76A, K81A, L139A, and D198A enhanced the expression of MenA, which increased MK production by 127.1%, 87.9%, 96.2%, 109.7% and 130.0% in wt-EmUbiA, respectively. In general, blocking UQ synthesis pathway for by site-directed mutagenesis of the active site of UbiA in E. meningoseptica was a promising strategy to increase MK production in E. meningoseptica. |
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| Keywords: | 4-hydroxybenzoate octaprenyl transferase (UbiA) Menaquinone Site-directed mutagenesis |
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